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利用纳米纤维支架培养的角膜缘和间充质干细胞治疗眼表损伤。

Treatment of ocular surface injuries by limbal and mesenchymal stem cells growing on nanofiber scaffolds.

机构信息

Institute of Molecular Genetics, Academy of Sciences, Prague, Czech Republic.

出版信息

Cell Transplant. 2010;19(10):1281-90. doi: 10.3727/096368910X509040. Epub 2010 Jun 23.

Abstract

Stem cell (SC) therapy represents a promising approach to treat a wide variety of injuries, inherited diseases, or acquired SC deficiencies. One of the major problems associated with SC therapy remains the absence of a suitable matrix for SC growth and transfer. We describe here the growth and metabolic characteristics of mouse limbal stem cells (LSCs) and mesenchymal stem cells (MSCs) growing on 3D nanofiber scaffolds fabricated from polyamide 6/12 (PA6/12). The nanofibers were prepared by the original needleless electrospun Nanospider technology, which enables to create nanofibers of defined diameter, porosity, and a basis weight. Copolymer PA6/12 was selected on the basis of the stability of its nanofibers in aqueous solutions, its biocompatibility, and its superior properties as a matrix for the growth of LSCs, MSCs, and corneal epithelial and endothelial cell lines. The morphology, growth properties, and viability of cells grown on PA6/12 nanofibers were comparable with those grown on plastic. LSCs labeled with the fluorescent dye PKH26 and grown on PA6/12 nanofibers were transferred onto the damaged ocular surface, where their seeding and survival were monitored. Cotransfer of LSCs with MSCs, which have immunosuppressive properties, significantly inhibited local inflammatory reactions and supported the healing process. The results thus show that nanofibers prepared from copolymer PA6/12 represent a convenient scaffold for growth of LSCs and MSCs and transfer to treat SC deficiencies and various ocular surface injuries.

摘要

干细胞(SC)疗法代表了一种有前途的方法,可以治疗多种损伤、遗传疾病或获得性 SC 缺乏症。与 SC 疗法相关的主要问题之一仍然是缺乏适合 SC 生长和转移的基质。我们在这里描述了在由聚酰胺 6/12(PA6/12)制成的 3D 纳米纤维支架上生长的小鼠角膜缘干细胞(LSCs)和间充质干细胞(MSCs)的生长和代谢特性。纳米纤维是通过原始的无针电纺 Nanospider 技术制备的,该技术能够制造出具有特定直径、孔隙率和基重的纳米纤维。选择共聚物 PA6/12 是基于其纳米纤维在水溶液中的稳定性、生物相容性以及作为 LSCs、MSCs 和角膜上皮细胞和内皮细胞系生长基质的优异性能。在 PA6/12 纳米纤维上生长的细胞的形态、生长特性和活力与在塑料上生长的细胞相当。用荧光染料 PKH26 标记并在 PA6/12 纳米纤维上生长的 LSCs 被转移到受损的眼表面,在那里监测它们的播种和存活情况。LSCs 与具有免疫抑制特性的 MSCs 的共转移显著抑制了局部炎症反应,并支持了愈合过程。因此,结果表明,由共聚物 PA6/12 制备的纳米纤维代表了 LSCs 和 MSCs 生长和转移的一种方便的支架,用于治疗 SC 缺乏症和各种眼表面损伤。

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